F/2 Culture Media

F/2 Medium Stock Solutions:

Quantity per L of Media

Compound

Stock Concentration

 
1.0 ml NaNO3 75.0 g/L dH2O Autoclave separately
1.0 ml NaH2PO4.H2O 5.0 g/L dH2O Autoclave separately
1.0 ml Na2SiO3.9H2O 30.0 g/L dH2O Autoclave separately
1.0 ml f/2 Trace Metal Solution (see recipe below) Filter sterilize
0.5 ml f/2 Vitamin Solution (see recipe below) Filter sterilize

Stock solutions (except vitamins) are considered 1000x. Seawater should be autoclaved and cooled prior to mixing the media.  We tend to sterile-filter artificial seawater rather than autoclaving to avoid reductions in dissolved carbon dioxide. A

Add all the f/2 components aseptically to your media once it has cooled to room temperature. For other algal groups (e.g. dinoflagellates, coccolithophores) use f/2-Si medium (prepare as for f/2 medium but omit Na2SiO3.9H2O). Sterile stock solutions can be stored at room temperature. Once opened, stock solutions should be stored at 4 °C.

Trace Metal Solution

To 950 mL distilled H2O add:

Quantity

Compound

Stock Solution

1.3 g FeCl3.6H2O  
8.7 g Na2EDTA.2H2O  
1.0 ml CuSO4.5H2O 980 mg / 100 ml dH20
1.0 ml Na2MoO4.2H2O 630 mg / 100 ml dH20
1.0 ml ZnSO4.7H2O 2.2 g / 100 ml dH20
1.0 ml CoCl2.6H2O 1.0 g / 100 ml dH20
1.0 ml MnCl2.4H2O 18.0 g / 100 ml dH20

Initially, the solution will be cloudy. Add 1N NaOH to adjust pH to about 4.5 to clear the solution. Bring final volume to one liter.

Vitamin Solution

To 950 mL dH2O add:

Quantity

Compound

1o Stock Solution

Tip

1.0 ml Vitamin B12 10 mg / 10 ml dH2O  
1.0 ml Biotin 10 mg / 10 ml dH2O Dissolve in a little 0.1N NaOH first
200.0 mg Thiamine HCl    

Make final volume up to 1.0 L with dH2O. Store primary stock solutions in freezer. Store completed vitamin solution in the refrigerator or freeze small aliquots.

 

References

Guillard R.R.L. and Ryther J.H. (1962) Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea Cleve. Can. J. Microbiol. 8: 229-239.

Guillard R.R.L. (1975) Culture of phytoplankton for feeding marine invertebrates. in “Culture of Marine Invertebrate Animals.” (eds: Smith W.L. and Chanley M.H.) Plenum Press, New York, USA. pp 26-60.